Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan ; Laboratory of Hepatocyte Differentiation, National Institute of Biomedical Innovation, Osaka 567-0085, Japan ; iPS Cell-Based Research Project on Hepatic Toxicity and Metabolism, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Stem Cell Reports. 2013 Oct 3;1(4):322-35. doi: 10.1016/j.stemcr.2013.08.006. eCollection 2013.
The establishment of self-renewing hepatoblast-like cells (HBCs) from human pluripotent stem cells (PSCs) would realize a stable supply of hepatocyte-like cells for medical applications. However, the functional characterization of human PSC-derived HBCs was not enough. To purify and expand human PSC-derived HBCs, human PSC-derived HBCs were cultured on dishes coated with various types of human recombinant laminins (LN). Human PSC-derived HBCs attached to human laminin-111 (LN111)-coated dish via integrin alpha 6 and beta 1 and were purified and expanded by culturing on the LN111-coated dish, but not by culturing on dishes coated with other laminin isoforms. By culturing on the LN111-coated dish, human PSC-derived HBCs were maintained for more than 3 months and had the ability to differentiate into both hepatocyte-like cells and cholangiocyte-like cells. These expandable human PSC-derived HBCs would be manageable tools for drug screening, experimental platforms to elucidate mechanisms of hepatoblasts, and cell sources for hepatic regenerative therapy.
从人类多能干细胞（PSCs）中建立具有自我更新能力的肝母细胞样细胞（HBCs）将实现用于医学应用的肝样细胞的稳定供应。然而，人类 PSC 衍生的 HBC 的功能特征还不够。为了纯化和扩增人 PSC 衍生的 HBC，将人 PSC 衍生的 HBC 培养在涂有各种类型的人重组层粘连蛋白（LN）的培养皿上。人 PSC 衍生的 HBC 通过整合素α 6 和β 1 附着在人层粘连蛋白 111（LN111）涂覆的培养皿上，并通过在 LN111 涂覆的培养皿上培养进行纯化和扩增，但不能在涂覆有其他层粘连蛋白同工型的培养皿上培养。在 LN111 涂覆的培养皿上培养，人 PSC 衍生的 HBC 可维持超过 3 个月，并具有分化为肝样细胞和胆管细胞样细胞的能力。这些可扩增的人 PSC 衍生的 HBC 将成为药物筛选的可控工具、阐明肝母细胞机制的实验平台以及肝再生治疗的细胞来源。
